基于DNA四面体HCR信号放大纳米体系的活细胞内circRNA检测与生物成像
PDF下载 (223)谢格格,陈静雯,刘浩蓉,喻盛容,唐科奇.基于DNA四面体HCR信号放大纳米体系的活细胞内circRNA检测与生物成像[J].宁波大学学报(理工版),2026,39(2):72-80.DOI:10.20098/j.cnki.1001-5132.2025.0518
XIE Gege,CHEN Jingwen,LIU Haorong,YU Shengrong,TANG Keqi.Detection and imaging of circRNA in living cells based on DNA tetrahedron HCR signal amplification nanosystem[J].Journal of Ningbo University(Natural Science & Engineering Edition),2026,39(2):72-80.DOI:10.20098/j.cnki.1001-5132.2025.0518
| Title: | Detection and imaging of circRNA in living cells based on DNA tetrahedron HCR signal amplification nanosystem |
| 作者: | 谢格格, 陈静雯, 刘浩蓉, 喻盛容, 唐科奇 |
| Author(s): | XIE Gege, CHEN Jingwen, LIU Haorong, YU Shengrong, TANG Keqi |
| 关键词: | DNA四面体; 荧光探针; 信号放大; 生物成像 |
| Keywords: | circRNA; DNAtetrahedron; fluorescence probe; signal amplification; biological imaging |
| 分类号: | R318 |
| DOI: | 10.20098/j.cnki.1001-5132.2025.0518 |
| 文献标识码: | A |
| 摘要: | 鉴于可靠生物标志物对早期诊断、治疗和预后评估疾病的关键作用，且现有研究证实circRNA具备成为潜在诊断与预后生物标志物的重要价值。本研究构建了一种基于DNA四面体的信号放大纳米体系（DTNH），该体系利用杂交链式反应（Hybridization Chain Reaction，HCR）进行信号放大，实现对活细胞中circRNA的检测及生物成像。HCR作为一种无酶、等温的信号放大技术，原理是通过目标物触发两个稳定的DNA发夹（H1和H2）发生交替、自持续的级联杂交反应，进而形成长双链DNA聚合物。基于此原理构建的DTNH由两种功能化的DNA四面体探针（DTNH-1和DTNH-2）构成。通过荧光法得出DTNH展现出优异的灵敏度和选择性，检测限为1.2×10–11 mol/L。同时，DTNH具有良好的生物相容性，凭借独特的纳米结构优势，能进入细胞，并成功用于活细胞内circMTO1的成像研究。开发的纳米体系不仅能动态监测细胞内circMTO1的表达水平，还可区分肿瘤细胞和正常细胞，同时为揭示circRNA空间表达特征提供了新方法，有望为开发精准高效的肿瘤诊断平台奠定一定基础。 |
| Abstract: | The discovery of reliable biomarkers is crucial for the early diagnosis, treatment, and prognostic assessment of diseases. Current research indicates that circRNA possesses significant potential as a diagnostic and prognostic biomarker. In this study, we developed a DNA tetrahedron-based signal amplification nanosystem (DTNH) that utilizes hybridization chain reaction (HCR) for signal amplification to detect circRNA in living cells and enable biological imaging. HCR is an enzyme-free, isothermal signaling amplification technique based on the principle of target-induced triggering of two stable DNA hairpins (H1 and H2), which then engage in alternating, self-sustaining cascade hybridization reactions to form long double-stranded DNA polymers. The DTNH constructed based on this principle comprises two types of functionalized DNA tetrahedron probes (DTNH-1 and DTNH-2). Fluorescence assays demonstrated that the DTNH exhibits excellent sensitivity and selectivity with a detection limit of 1.2×10–11 mol/L. Additionally, the DTNH showcases good biocompatibility; due to its unique nanoscale structural advantages, it can enter cells effectively and has been successfully applied for imaging studies of circMTO1 within live cells. The developed nanoplatform can not only dynamically monitor the expression levels of circMTO1 within cells but also distinguish between tumor cells and normal cells. At the same time, it provides a new method for revealing the spatial expression characteristics of circRNA, which is expected to lay a certain foundation for developing precise and efficient tumor diagnostic platforms. |
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| 备注/Memo: | 收稿日期：2025-05-19 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ 基金项目：国家自然科学基金（U20A20121） 第一作者：谢格格，硕士研究生，主要研究方向为化学生物传感。E-mail: 17729177520@163.com *通信作者：唐科奇，教授，主要研究方向为质谱学理论及技术应用。E-mail: tangkeqi@nbu.edu.cn 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ |
